頁籤選單縮合
題名 | Molecular Characterization of a Plasmid-Borne (pTE31) Erythromycin Resistance Determinant (erm-31) from Lactobacillus reuteri AE72=洛德乳酸桿菌AE72株抗藥質體pTE31之抗紅黴素基因erm-31的分子特性 |
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作者 | 林春福; 方榮華; 許添桓; 楊忠亮; 許慶宗; 張登欽; | 書刊名 | 中華民國獸醫學會雜誌 |
卷期 | 25:2 1999.06[民88.06] |
頁次 | 頁88-95 |
分類號 | 437.243 |
關鍵詞 | 洛德乳酸桿菌; Lactobacillus reuteri; rRNA methylase gene; |
語文 | 英文(English) |
中文摘要 | 洛德乳酸桿菌(Lactobacillus reuteri) AE72株,其抗紅黴素(erythromycin)的 最小抑制濃度為 > 516 ug/mL。 本菌株經由翦除 (curing)、電孔 (electroporation)、雜 合 (hybridization) 和選殖 (cloning) 等實驗分析,證實含有一 4.4 Kb 抗紅黴素內源性 質體 pTE31, 其抗紅黴素基因 erm-31( 含於 1.2Kb Hindlll- Pstl 片段中 ) 也成功地選 殖於 E. coli TG1 勝任細胞 (competent cell)。erm-31 經 DNA 定序,結果顯示具有一個 可轉譯出 250 個氨基酸之開讀框 (open reading frame; ORF), 屬於 rRNA methylase gene, 此基因長度為 750 bp, 與 Streptococcus faecalis Tn917 之 erm 基因 (735bp) 比對,在核酸及氨基酸各具有 93 %及 90 %之相似性。為測定 erm-31 基因之活性,將帶 有 erm-31 基因之重組質體 pUE3112 進行 maxicell 分析,其蛋白產物經電泳後, 分子量 為 27.5 kDa, 此與 DNA 序列之預測值相同。 本實驗為首次在洛德乳酸桿菌中,發現屬於 erm AM type 抗紅黴素基因 DNA 序列者。 |
英文摘要 | This study demonstrates that Lactobacillus reuteri AE72 is resistant to erythromycin (Em') with a minimum inhibitory concentration (MIC) of 516 ug/mL. Through a series of analyses including curing, electroporation, hybridization, and cloning experimentation, a 4.4 kb indigenous plasmid (pTE31) of L. reuteri AE72 was successfully identified as Em resistant (Em). A 1.2 kb Hindlll-Pstl DNA fragment containing the Em' determinant (erm-31) from pTE31 was also. successfully cloned into E. coli TG1. Determining the nucleotide sequence of erm-31 revealed an open reading frame (ORF) for a putative 250-amino acid rRNA methylase gene (erm). This structural erm gene, 750 bp in length, was highly related (ca. 93% nucleotide and ca. 90% amino acid identity) to the 735 bp erm gene of the Tn917 of Streptococcus faecalis, A recombinant plasmid pUE3112 containing the erm-31 determinant from pTE31 was subjected to maxicell analysis to determine the activity of the putative erm-31 gene. The observed molecular mass of the synthesized protein, based on electrophoretic mobility, correlated with the 27.5 kDa protein predicted from the DNA sequence. To our knowledge, this work characterizes erm AM type of Em' determinant from L. reuteri to be to the nucleotide sequence level for the first time. |
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